RESUMO
In this chapter, a protocol to design affinity chromatography matrices with short peptide ligands immobilized for protein purification is described. The first step consists of the synthesis of a combinatorial peptide library on the hydroxymethylbenzoyl (HMBA)-ChemMatrix resin by the divide-couple-recombine (DCR) method using the Fmoc chemistry. Next, the library is screened with the protein of interest labeled with a fluorescent dye or biotin. Subsequently, peptides contained on positive beads are identified by tandem matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS), and those sequences showing greater consensus are synthesized in larger quantities and immobilized on chromatographic supports. Finally, target protein adsorption on peptide affinity matrices is evaluated through equilibrium adsorption isotherms and breakthrough curves.
Assuntos
Cromatografia de Afinidade , Técnicas de Química Combinatória , Biblioteca de Peptídeos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Fukui functions (FFs) are chemical descriptors that are useful to explain the reactivity of systems toward electron transfer. Whereas they have been widely employed for molecules, their application to extended systems is scarce. One of the reasons for the limited development of such analysis in solids is the improper evaluation of FFs in the usual computational approaches based on density functional theory and periodic boundary conditions. In this work we compare the available approaches and propose a new method based on the interpolation of partially charged systems that mitigates some of the problems encountered. We discuss the reactivity of alkaline earth oxides (MgO, CaO, SrO, and BaO) in terms of the FF analysis, providing a robust way to account for the higher reactivity of surface oxygen sites compared with bulk sites.
RESUMO
The aim of the study was to evaluate the effects of a cashew nut shell oil and commercial castor oil blend (CNSL-Castor oil) on the performance and microbiota of broiler chickens with and without coccidiosis challenge. A total of 864 one-day-old male chicks (Cobb) were randomly distributed to receive 6 treatments (8 pens/treatment; 18 chicks/pen) in a 3 × 2 factorial, with 3 additives (control [non-additives], 100 ppm sodium monensin, or 0.15% CNSL-Castor oil blend), and 2 levels of coccidiosis challenge at 14 D of age (unchallenged or inoculated by gavage with 1 mL of solution containing oocysts sporulated with Eimeria tenella, Eimeria acervulina, and Eimeria maxima). No differences in productive performance were observed among treatments in the pre-challenge period and in unchallenged birds (P > 0.05). Seven-days post-challenge, birds receiving monensin performed better than birds in the positive control group (non-additive and challenge) or in the CNSL-Castor oil group (P > 0.05). However, 14 D post-challenge, birds supplemented with CNSL-Castor oil presented higher weight gain and better feed conversion (P > 0.05), without any change in feed intake (P > 0.05). During the accumulated period (1 to 42 D of age), the live weight, weight gain, and feed intake did not differ between the CNSL-Castor oil and monensin groups, both of which presented higher values than the positive control. Lactobacillus spp. and Clostridium perfringens numbers were increased in the challenged birds (P < 0.05). CNSL-Castor oil supplementation reduced Clostridium cluster XIV, C. perfringens, and S. aureus, compared with the monensin and control groups (P > 0.05). In addition, the CNSL-Castor oil group presented the highest number of Lactobacillus spp. copies, followed by the monensin and positive control groups (P > 0.05). Thus, monensin and CNSL-Castor oil effectively minimized the impact of coccidiosis at different times. While monensin acts as an antimicrobial, CNSL-Castor oil modulates the intestinal microbiota with antimicrobial action against gram-positive bacteria, mainly C. perfringens and S. aureus.
Assuntos
Anti-Infecciosos/farmacologia , Galinhas/imunologia , Microbioma Gastrointestinal/efeitos dos fármacos , Monensin/farmacologia , Óleos de Plantas/farmacologia , Anacardium/química , Ração Animal/análise , Animais , Anti-Infecciosos/classificação , Óleo de Rícino/farmacologia , Galinhas/crescimento & desenvolvimento , Galinhas/microbiologia , Galinhas/fisiologia , Coccidiose/imunologia , Coccidiose/veterinária , Dieta/veterinária , Suplementos Nutricionais/análise , Eimeria/fisiologia , Masculino , Doenças das Aves Domésticas/imunologia , Distribuição AleatóriaRESUMO
Infection with Eimeria sp. results in the activation of multiple facets of the host immune system; the use of phytogenics can modulate the inflammatory response and improve the performance of the challenged animal. The aim of this study was to evaluate the effect of a commercial blend of cashew nut shell liquid (CNSL) and castor oil on the immune response of broilers challenged with coccidiosis. A total of 864 one-day-old male chicks (Cobb 500) were randomly distributed into six treatment groups (8 pens/treatment and 18 chicks/pen) in a three-by-two factorial design with three additives: control (non-additive), 100 ppm of monensin or 0.15% CNSL-castor oil. Challenge status was determined twice at 14 days of age. Unchallenged birds were inoculated by gavage with oocysts sporulated with Eimeria tenella, Eimeria acervulina and Eimeria maxima. Although the positive control (non-additive and challenged) and CNSL-castor oil treatment groups exhibited similar variation in weight gain (ΔBWG) compared to unchallenged birds fed without additives, the variation observed in birds fed diets containing CNSL-castor oil was associated with a higher maintenance requirement and not feed efficiency. In the second week after infection, ΔBWG of the CNSL-castor oil treatment group did not significantly change compared to the other treatment groups. At days 7 and 14 post-challenge, there was a higher excretion of oocysts in the control group, whereas the CNSL-castor oil and monensin groups did not differ. The CNSL-castor oil group exhibited increased gene expression of interferon (IFN), interleukin 6 (IL-6) and tumor necrosis factor (TNF), while the control group exhibited increased expression of cyclooxygenase (COX) and IL-1. The heterophils/lymphocyte ratio was low for the monensin treatment group. The unchallenged birds that received monensin treatment presented higher gene expression of IFN, COX and IL-1 compared to the other treatments, while the CNSL-castor oil group exhibited reduced gene expression, except for TNF. The commercial blend of cashew nut liquid and castor oil modulated the inflammatory response against Eimeria spp. In the absence of the parasite, there was no stimulation of genes involved in the inflammatory response, demonstrating that the blend is an effective tool in specifically modulating the immune system of birds afflicted with coccidiosis.
Assuntos
Anacardium/química , Galinhas/imunologia , Coccidiose/imunologia , Eimeria/fisiologia , Óleos de Plantas/administração & dosagem , Doenças das Aves Domésticas/imunologia , Ração Animal , Animais , Galinhas/parasitologia , Coccidiose/parasitologia , Dieta/veterinária , Masculino , Nozes/química , Doenças das Aves Domésticas/parasitologia , Distribuição Aleatória , Aumento de PesoRESUMO
The experiment was conducted to study the effect of pure glycerin supplementation (GLYC) in the drinking water of broilers subjected to heat stress and feed restriction. Water with 0, 1, or 2% glycerin was provided ad libitum to broilers in six hours of feed restriction. The birds were housed in two environments: thermoneutral (TN) - 25 ° C, and cyclic heat stress (HS) - 12h with 250C, 6h with 320C, 3h of 250 to 320C and 3h of 320 to 250C. The experimental design was randomized in a factorial arrangement of 2 x 3 + 2, with three GLYC levels, two environments and one control group with ad libitum feeding each environment. When submitted to HS, broiler receiving 2% glycerin presented higher weight gain (WG), water consumption (WC), feed intake (FI) and energy consumption (EI) than those in the other treatments, contrarily to birds in TN, where increasing GLYC levels decreased those responses. Broilers submitted to feed restriction presented reduced FI, but better feed conversion (FCR), independently of rearing environment or GLYC levels. Broilers under HS submitted to feed restriction and receiving 2% GLYC presented higher WC (p < 0.05) and similar WG as the controls, differently from the restricted-fed broilers under TN receiving 2% GLYC, which WC and WG were lower than the controls. The inclusion of 2% pure glycerin in the drinking water may compensate the negative effects on performance caused by feed restriction in broilers submitted to heat stress.(AU)
Assuntos
Animais , Glicerol/análise , Galinhas/crescimento & desenvolvimento , Restrição Calórica/veterinária , Transtornos de Estresse por Calor/veterinária , Ingestão de Líquidos , Aditivos Alimentares/análise , Fenômenos Fisiológicos da Nutrição Animal , Aumento de Peso , Ingestão de Alimentos , Água Potável/análiseRESUMO
The experiment was conducted to study the effect of pure glycerin supplementation (GLYC) in the drinking water of broilers subjected to heat stress and feed restriction. Water with 0, 1, or 2% glycerin was provided ad libitum to broilers in six hours of feed restriction. The birds were housed in two environments: thermoneutral (TN) - 25 ° C, and cyclic heat stress (HS) - 12h with 250C, 6h with 320C, 3h of 250 to 320C and 3h of 320 to 250C. The experimental design was randomized in a factorial arrangement of 2 x 3 + 2, with three GLYC levels, two environments and one control group with ad libitum feeding each environment. When submitted to HS, broiler receiving 2% glycerin presented higher weight gain (WG), water consumption (WC), feed intake (FI) and energy consumption (EI) than those in the other treatments, contrarily to birds in TN, where increasing GLYC levels decreased those responses. Broilers submitted to feed restriction presented reduced FI, but better feed conversion (FCR), independently of rearing environment or GLYC levels. Broilers under HS submitted to feed restriction and receiving 2% GLYC presented higher WC (p < 0.05) and similar WG as the controls, differently from the restricted-fed broilers under TN receiving 2% GLYC, which WC and WG were lower than the controls. The inclusion of 2% pure glycerin in the drinking water may compensate the negative effects on performance caused by feed restriction in broilers submitted to heat stress.
Assuntos
Animais , Galinhas/crescimento & desenvolvimento , Glicerol/análise , Ingestão de Líquidos , Restrição Calórica/veterinária , Transtornos de Estresse por Calor/veterinária , Aditivos Alimentares/análise , Aumento de Peso , Fenômenos Fisiológicos da Nutrição Animal , Ingestão de Alimentos , Água Potável/análiseRESUMO
We isolated and identified functional groups of bacteria in the rumen of Creole goats involved in ruminal fermentation of native forage shrubs. The functional bacterial groups were evaluated by comparing the total viable, total anaerobic, cellulolytic, hemicellulolytic, and amylolytic bacterial counts in the samples taken from fistulated goats fed native forage diet (Atriplex lampa and Prosopis flexuosa). Alfalfa hay and corn were used as control diet. The roll tubes method increased the possibility of isolating and 16S rDNA gene sequencing allowed definitive identification of bacterial species involved in the ruminal fermentation. The starch and fiber contents of the diets influenced the number of total anaerobic bacteria and fibrolytic and amylolytic functional groups. Pseudobutyrivibrio ruminis and Pseudobutyrivibrio xylanivorans were the main species isolated and identified. The identification of bacterial strains involved in the rumen fermentation helps to explain the ability of these animals to digest fiber plant cell wall contained in native forage species.
Assuntos
Ração Animal , Dieta/métodos , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Rúmen/microbiologia , Animais , Carga Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Cabras , Bactérias Gram-Positivas/genética , Bactérias Gram-Positivas/metabolismo , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Objetivo. Estimar heredabilidades, correlaciones fenotípicas y genotípicas entre 24 características de tipo y producción de leche en vacas Holstein del departamento de Antioquia. Materiales y métodos. Se analizaron un total de 2395 registros de vacas Holstein. Se estimaron componentes de varianza mediante el procedimiento de máxima verosimilitud restricta con modelos animales bi-característicos. Para las características de tipo, el modelo incluyó los efectos fijos de finca, año de nacimiento y país de origen del padre de la vaca, el grupo contemporáneo (año-mes-clasificador) y la covariable edad a la clasificación. Para la producción de leche el modelo incluyó los efectos fijos de finca, año de parto, país y año de nacimiento del padre. Resultados. Las heredabilidades encontradas para las características de tipo variaron de 0.04 a 0.44 y para producción de leche fue de 0.22, los errores estándar de la heredabilidad fueron inferiores a 0.06. Las correlaciones genéticas de producción de leche con las características profundidad de la ubre (-0.72) fue alta, mientras las correlaciones más altas entre las características de tipo fueron entre extremo anterior y tamaño (0.90), ángulo de la pezuña y profundidad del talón (0.98), profundidad del talón y uniformidad de pezuñas (0.85); y ligamento suspensorio medio y colocación de pezones posteriores (0.76). Conclusiónes. Se encontró una considerable variación genética en características de tipo dentro de la población Holstein de Antioquia. Las correlaciones genéticas entre algunas características de tipo fueron altas e indican la posibilidad de reducir el número de características evaluadas en cada animal.
Assuntos
Animais , Bovinos , Genética , LeiteRESUMO
Los modelos de evaluación genética multirracial permiten la obtención de los valores genéticos de animales perteneciente a una población de diferente composición racial. La estructura de este tipo de modelo va a depender de la característica a evaluar y de la población multirracial a ser analizada, obteniendose al final de la evaluación valores genéticos aditivos y no aditivos, los cuales permiten tomar decisiones y direccionar los apareamientos de una forma más acertada.
Assuntos
Bovinos , Bovinos , DesmameRESUMO
Um experimento foi realizado para avaliar o efeito da adição de extratos cítricos e ractopamina a dietas de suínos em terminação. Foram utilizados 108 suínos (54 machos e 54 fêmeas), homogêneos geneticamente e peso vivo médio inicial de 61 quilogramas. O delineamento experimental foi de blocos ao acaso, em arranjo fatorial 3x3 (três níveis de ractopamina (RAC): 0, 10 e 20 ppm e três níveis de extratos cítricos (EC): 0, 250 e 500 ppm) com quatro repetições. Foram avaliadas amostras do músculo longissimus dorsi quanto à umidade, cinzas, proteínas, lipídios e perfil de ácidos graxos. Os teores de proteína para a inclusão de 20 ppm de RAC foram em média 5,5% superiores (P 0,05) aos dois níveis de EC na dieta. A umidade do músculo nas amostras dos animais que receberam 500 ppm de EC e 20 ppm de RAC foi 4,3% superior (P 0,05) ao controle e 500 ppm de extratos cítricos. Os teores do ácido linoléico da interação 500 ppm de EC e 10 ppm de RAC foi 18% superior (P 0,05) em relação à inclusão de 500 ppm de extratos cítricos. Os teores do ácido -Linolênico do controle foi 33,5% superior (P 0,05) aos níveis de extratos cítricos, ractopamina e suas interações. A concentração do ácido araquidônico da interação 250 ppm de EC e 20 ppm de RAC foi 36% superior (P 0,05) aos teores de 20 ppm de ractopamina. Níveis mais altos de ractopamina às dietas influenciam os teores de
RESUMO
Um experimento foi realizado para avaliar o efeito da adição de extratos cítricos e ractopamina a dietas de suínos em terminação. Foram utilizados 108 suínos (54 machos e 54 fêmeas), homogêneos geneticamente e peso vivo médio inicial de 61 quilogramas. O delineamento experimental foi o inteiramente casualizado, bloqueado por sexo e com nove tratamentos: T1. controle (C) (0 ppm de ractopamina e 0 ppm de extratos cítricos), T2. C + 10 RAC (ractopamina, em ppm), T3. C + 20 RAC, T4. C + 250 EC (extratos cítricos, em ppm), T5. C+ 500 EC, T6. C + 250 EC + 10 RAC, T7. C + 250 EC + 20 RAC, T8. C+ 500 EC + 10 RAC e T9. C + 500 EC + 20 RAC. Foram utilizados dois sexos, com duas repetições e seis animais por unidade experimental. Foram avaliadas amostras do músculo Longissimus dorsi quanto à umidade, cinzas, proteínas, lipídios e perfil de ácidos graxos. Os teores de proteína para a inclusão de 20 ppm de RAC foram em média 5,5% superiores (P<0,05) aos dois níveis de EC na dieta. A umidade do músculo nas amostras dos animais que receberam 500 ppm de EC e 20 ppm de RAC foi 4,3% superior (P<0.05) ao controle e 500 ppm de extratos cítricos. Os teores do ácido linoléico da interação 500 ppm de EC e 10 ppm de RAC foi 18% superior (P<0,05) em relação à inclusão de 500 ppm de extratos cítricos. Os teores do ácido a-Linolênico do controle foi 33,5% superior (P<0,05) aos níveis de extratos cítricos, ractopamina e suas interações. A concentração do ácido araquidônico da interação 250 ppm de EC e 20 ppm de RAC foi 36% superior (P<0,05) aos teores de 20 ppm de ractopamina. Níveis mais altos de ractopamina às dietas influenciam os teores de proteína e umidade do músculo. Os extratos cítricos influenciam os teores do ácido graxo láurico. A adição de ractopamina altera o perfil de alguns ácidos graxos insaturados do músculo Longissimus dorsi.
This study was carried out to evaluate the effect of the addition of the citrus extracts and ractopamine in finishing pig diets. Hundred eight pigs were used (54 males and 54 females) in a completely randomized design, blocked by sex and distributed in nine treatments: T1. control (C) (0 ppm of the ractopamine e 0 ppm of the citrus extracts), T2. C + 10 RAC (ractopamine, ppm), T3. C + 20 RAC, T4. C + 250 EC (citrus extracts, ppm), T5. C+ 500 EC, T6. C + 250 EC + 10 RAC, T7. C+ 250 EC + 20 RAC, T8. C+ 500 EC + 10 RAC e T9. C+ 500 EC + 20 RAC. We used two sexes, with two replications and six animals per experimental unit. The levels of protein for the inclusion of 20 ppm of RAC were on average 5.5% higher (P <0.05) the two EC levels in the diet. The moisture in the muscle samples from the animals that received 500 ppm EC and 20 ppm RAC was 4.3% higher (P <0.05) the control and 500 ppm of citrus extracts. The levels of linoleic acid in the interaction of 500 ppm EC and 10 ppm RAC was 18% higher (P <0.05) compared to the inclusion of 500 ppm of citrus extracts. The levels of a-linolenic acid of the control was 33.5% higher (P <0.05) of citrus extracts levels, ractopamine and their interactions. The concentration of arachidonic acid from the interaction of EC 250 ppm and 20 ppm RAC was 36% higher (P <0.05) to levels of 20 ppm of ractopamine. Higher levels of ractopamine in the diet influence the levels of protein and moisture of the muscle. Citrus extracts influence the levels of the fatty acid lauric acid. The addition of ractopamine change the profile of some unsaturated fatty acids of the Longissimus dorsi.
Assuntos
Animais , Ácido Ascórbico/administração & dosagem , Suínos , Flavonoides/administração & dosagem , Agonistas Adrenérgicos beta , Músculos do Dorso/química , Carne/análiseRESUMO
This study was conducted to evaluate if supplementing bypass fat to cows under silvopastoral systems, increases the concentration of unsaturated fatty acids in milk, thus improving the saturated/ unsaturated ratio without a negative effect on total milk yield in fat or protein. Two concentrations of two different sources of bypass fat were evaluated for 40 days, each in a group of 24 multiparous Lucerna (Colombian breed) cows. A cross-over design of 8 Latin squares 3 x 3 was used. The variables submitted to analysis were body condition, daily milk production and milk composition. Body condition, milk yield and milk quality were not different but there was a significant decrease in the amount of saturated fatty acid in both experiments while the unsaturated fat increased significantly in experiment 1 and remained stable in experiment 2. Results, such as these have as far as we know, not been reported previously and they provide an approach for the improvement of milk as a "functional food".
Assuntos
Bovinos/fisiologia , Gorduras na Dieta/administração & dosagem , Ácidos Graxos Insaturados/análise , Ácidos Graxos/análise , Lactação/metabolismo , Proteínas do Leite/análise , Leite/química , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bovinos/metabolismo , Estudos Cross-Over , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Feminino , Distribuição AleatóriaRESUMO
Objetivo. Estimar la heredabilidad del intervalo de partos en bovinos cruzados. Materiales y métodos. Se utilizó información de la base de datos de la hacienda La Leyenda, ubicada en el municipio de Caucasia (Antioquia). Se realizó un análisis uni-característico empleando el modelo animal. En el modelo se consideraron los efectos fijos del grupo genético, año de nacimiento, época de nacimiento y el número de partos y como efectos aleatorios, el efecto aditivo directo, de ambiente permanente y residual. Resultados. Se obtuvo una heredabilidad de 0.15 ± 0.07 y una media fenotípica de 544 ± 97 días. Conclusión. La heredabilidad para el intervalo de partos fue baja, por lo que se puede lograr poco progreso genético seleccionando para esta característica.
Assuntos
Bovinos , Animais , Lactente , Bovinos , Carne , Parto , Parto/genética , Reprodução , Bovinos/crescimento & desenvolvimento , Bovinos/genética , Carne/análise , Carne/microbiologia , Reprodução/genéticaRESUMO
The adhE gene of Escherichia coli encodes a multifunctional ethanol oxidoreductase (AdhE) that catalyzes successive reductions of acetyl-CoA to acetaldehyde and then to ethanol reversibly at the expense of NADH. Mutant JE52, serially selected for acquired and improved ability to grow aerobically on ethanol, synthesized an AdhE(A267T/E568K) with two amino acid substitutions that sequentially conferred improved catalytic properties and stability. Here we show that the aerobic growth ability on ethanol depends also on protection of the mutant AdhE against metal-catalyzed oxidation by the chaperone DnaK (a member of the Hsp70 family). No DnaK protection of the enzyme is evident during anaerobic growth on glucose. Synthesis of DnaK also protected E. coli from H2O2 killing under conditions when functional AdhE is not required. Our results therefore suggest that, in addition to the known role of protecting cells against heat stress, DnaK also protects numerous kinds of proteins from oxidative damage.
Assuntos
Álcool Desidrogenase/metabolismo , Aldeído Oxirredutases/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Escherichia coli , Escherichia coli/metabolismo , Proteínas de Choque Térmico HSP70/fisiologia , Complexos Multienzimáticos/metabolismo , Estresse Oxidativo , Aerobiose , Anaerobiose , Escherichia coli/crescimento & desenvolvimento , Etanol/farmacologia , Mutação , OxirreduçãoRESUMO
We report a boy in whom and advanced Hodgkin's disease, nodular sclerosis variety, was diagnosed at 5 years of age and treated with exclusive chemotherapy. After 16 years of remission, he presented with a relapse of the disease, with a different histological pattern and was subjected to chemotherapy (C-MOPP/AVB) and unilateral axillary irradiation, obtaining a complete remission of the disease. Four months later, the patient is asymptomatic and without evidences of relapse.
Assuntos
Doença de Hodgkin/terapia , Recidiva Local de Neoplasia/terapia , Pré-Escolar , Humanos , Masculino , Indução de RemissãoRESUMO
We report a boy in whom an advanced Hodgkin's disease, nodular sclerosis variety was diagnosed at 5 yeras of age and treated with exclusive chemotherapy. After 16 years of remission, he presented with a relapsed of the disease with a different histological pattern and was subjected to chemotherapy (C-MOPP/AVB) and unilateral axillary irradiation, obtaining a complete remission of the disease. Four months later, the patients is asymptomatic and without evidences of relapse
Assuntos
Humanos , Masculino , Adulto , Doença de Hodgkin/tratamento farmacológico , Recidiva Local de Neoplasia/patologia , Esquema de Medicação , PrognósticoRESUMO
A clinical, biochemical, and pathological study was performed in 38 chronic HBsAg carriers. The study group is a part of 393 carriers found among 117 705 voluntary blood donors at the National Blood Bank, Hospital del Salvador, Santiago, Chile. None of the 38 carriers had a past history of illicit drug abuse, hepatitis, or work involving a high risk of hepatitis B virus infection. Ten individuals had a normal liver biopsy, 17 reactive non-specific hepatitis, one fatty changes, four chr onic persistent hepatitis, one aggressive hepatitis, two post-necrotic cirrhosis, and three alcoholic cirrhosis. There was not a close correlation between liver function test and liver histology. The most significant laboratory finding was the postivity of alpha fetoprotein in two cases. During the follow-up the two alpha fetoprotein patients presented a hepatocarcinoma 12 and 14 months after admission to the study.